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Sequence-specific DNA uptake in Haemophilus transformation.

K L Sisco, H O Smith

    Proceedings of the National Academy of Sciences of the United States of America
    |February 1, 1979
    PubMed
    Summary
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    Haemophilus bacteria specifically absorb their own DNA using unique uptake sites. Researchers identified two small DNA fragments containing these recognition sequences, crucial for bacterial transformation.

    Area of Science:

    • Microbiology
    • Molecular Biology
    • Genetics

    Background:

    • Haemophilus species exhibit selective DNA uptake during transformation.
    • Understanding the mechanism of this DNA specificity is crucial for genetic studies.

    Purpose of the Study:

    • To investigate the sequence-specific DNA uptake mechanism in Haemophilus.
    • To identify the specific DNA sequences responsible for recognition and uptake.

    Main Methods:

    • Cloning of an 8.1-kilobase (kb) H. parainfluenzae DNA fragment in E. coli.
    • Using 5'32P end-labeled DNA fragments to assay uptake by competent Haemophilus cells.
    • Restriction endonuclease mapping to identify DNA fragments with uptake activity.

    Main Results:

    Related Experiment Videos

    • Competent Haemophilus cells specifically absorbed the 8.1-kb H. parainfluenzae DNA fragment, but not the E. coli vector DNA.
    • Two small fragments (120 and 140 base pairs) within the 8.1-kb fragment retained DNA uptake ability.
    • These fragments are located 3.8 kb apart and likely contain short (8-12 bp) common recognition sequences.
    • DNA competition assays indicated approximately 600 copies of these uptake sites per Haemophilus genome.

    Conclusions:

    • Haemophilus DNA uptake is mediated by specific recognition sites on the DNA molecule.
    • These sites are present in approximately 600 copies within the Haemophilus genome.
    • The identified fragments provide a basis for further understanding bacterial transformation specificity.